發(fā)布時(shí)間：2018-11-16 16:53

【摘要】：目的通過(guò)大鼠腹腔注射硝酸甘油(GTN)建立急性偏頭痛模型。造模成功后使用Western blot技術(shù)檢測(cè)大鼠三叉神經(jīng)節(jié)上P2X7受體和c-fos的表達(dá)變化;使用免疫熒光檢測(cè)衛(wèi)星膠質(zhì)細(xì)胞上P2X7受體和膠質(zhì)纖維酸性蛋白(GFAP)的表達(dá)情況。最后通過(guò)腹腔注射P2X7受體抑制劑亮藍(lán)(BBG)觀察其對(duì)三叉神經(jīng)脊束核c-fos和降鈣素基因相關(guān)肽(CGRP)表達(dá)的影響,來(lái)探究P2X7受體是否參與偏頭痛的發(fā)病及其可能的機(jī)制。材料和方法1.Sprague-Dawley大鼠10只,隨機(jī)分為2組:腹腔注射生理鹽水組(n=5,2ml/Kg)和腹腔注射GTN組(n=5,10mg/Kg)。給藥前使用Von Frey測(cè)痛儀測(cè)量各組大鼠后爪上的基礎(chǔ)痛覺(jué)閾值,之后分別在給藥第1、2和4小時(shí)時(shí)再次測(cè)量大鼠后爪上的痛覺(jué)閾值。使用免疫熒光觀察大鼠三叉神經(jīng)節(jié)P2X7受體和GFAP的表達(dá)。2.Sprague-Dawley大鼠20只,隨機(jī)分為4組:正常組(n=5)和腹腔注射GTN 1小時(shí)組(n=5,10mg/Kg)、腹腔注射GTN 2小時(shí)組(n=5,10mg/Kg)和腹腔注射GTN 4小時(shí)組(n=5,10mg/Kg)。Western blot技術(shù)檢測(cè)大鼠三叉神經(jīng)節(jié)上P2X7受體和c-fos的表達(dá)變化。3.Sprague-Dawley大鼠20只,隨機(jī)分為4組:腹腔注射生理鹽水2小時(shí)組(n=5,2ml/Kg)、腹腔注射GTN 2小時(shí)組(n=5,10mg/Kg)、腹腔注射GTN+生理鹽水2小時(shí)組(n=5,10mg/Kg,2ml/Kg)和腹腔注射GTN+BBG 2小時(shí)組(n=5,10mg/Kg)。使用Western blot技術(shù)檢測(cè)大鼠三叉神經(jīng)脊束核c-fos和CGRP的表達(dá)。結(jié)果1.痛覺(jué)閾值:腹腔注射GTN后,大鼠后爪上的痛覺(jué)閾值開(kāi)始下降,在第2小時(shí)下降最為明顯,明顯低于生理鹽水組(p0.05)。2.免疫熒光:三叉神經(jīng)節(jié)上的P2X7受體和GFAP共表達(dá)。3.Western blot:腹腔注射GTN后,大鼠衛(wèi)星膠質(zhì)細(xì)胞P2X7受體表達(dá)開(kāi)始下降,在第2小時(shí)下降最為明顯,明顯低于正常組(p0.05)。而三叉神經(jīng)節(jié)和三叉神經(jīng)脊束核c-fos和CGRP的表達(dá)明顯高于正常組(p0.05)。在使用P2X7受體抑制劑BBG后c-fos和CGRP增加的更加明顯(p0.05)。結(jié)論1.P2X7受體特異性表達(dá)在大鼠三叉神經(jīng)節(jié)衛(wèi)星膠質(zhì)細(xì)胞上。2.急性偏頭痛大鼠模型三叉神經(jīng)節(jié)P2X7受體表達(dá)降低,提示P2X7受體在偏頭痛中可能有一定作用。3.急性偏頭痛大鼠腹腔注射BBG后,三叉神經(jīng)脊束核c-fos和CGRP的表達(dá)進(jìn)一步增高,表明衛(wèi)星膠質(zhì)細(xì)胞P2X7受體在偏頭痛中可能具有保護(hù)作用。
[Abstract]:Objective to establish acute migraine model by intraperitoneal injection of nitroglycerin (GTN) in rats. Western blot technique was used to detect the expression of P2X7 receptor and c-fos in rat trigeminal ganglion and the expression of P2X7 receptor and glial fibrillary acidic protein (GFAP) in satellite glial cells was detected by immunofluorescence. Finally, the effects of P2X7 receptor inhibitor, brilliant blue (BBG), on the expression of c-fos and calcitonin gene-related peptide (CGRP) in trigeminal spinal tract nucleus were observed in order to explore whether P2X7 receptor is involved in the pathogenesis of migraine and its possible mechanism. Materials and methods Ten 1.Sprague-Dawley rats were randomly divided into two groups: saline group (n = 5) and GTN group (n = 10 mg / kg). The basic pain threshold on the posterior paw of rats in each group was measured by Von Frey before administration, and the pain threshold on the posterior paw of rats was measured again at the 2nd and 4th hour after administration of the drug. The expression of P2X7 receptor and GFAP in trigeminal ganglion was observed by immunofluorescence. Twenty 2.Sprague-Dawley rats were randomly divided into 4 groups: normal group (nong5) and intraperitoneal injection of GTN (10 mg / kg). The expression of P2X7 receptor and c-fos in the trigeminal ganglion was detected by intraperitoneal injection of GTN (10 mg / kg) and intraperitoneal injection of GTN (10 mg / kg). Western blot) in 20 3.Sprague-Dawley rats. They were randomly divided into 4 groups: intraperitoneal injection of normal saline for 2 hours group (n = 5), intraperitoneal injection of GTN for 2 hours group (n = 5), intraperitoneal injection of GTN saline group for 2 hours (n = 10 mg / kg), intraperitoneal injection of GTN group for 2 hours (n = 5). 2ml/Kg and intraperitoneal injection of GTN BBG for 2 hours (n = 5 10 mg / kg). Western blot technique was used to detect the expression of c-fos and CGRP in the spinal trigeminal nucleus of rats. Result 1. Pain threshold: after intraperitoneal injection of GTN, the pain threshold on the posterior claw of rats began to decrease, which was the most obvious at the 2nd hour, which was significantly lower than that in the saline group (p0.05). Immunofluorescence: P2X7 receptor and GFAP co-expressed in trigeminal ganglion. After 3.Western blot: was injected intraperitoneally with GTN, the expression of P2X7 receptor in rat satellite glial cells began to decrease, and at the 2nd hour, the expression of P2X7 receptor was significantly lower than that in normal group (p0.05). The expression of c-fos and CGRP in trigeminal ganglion and spinal trigeminal nucleus was significantly higher than that in normal group (p0.05). C-fos and CGRP increased more significantly after the use of P2X7 receptor inhibitor BBG (p0.05). Conclusion 1.P2X7 receptor is specifically expressed in rat trigeminal ganglion satellite glial cells. 2. 2. The expression of P2X7 receptor in trigeminal ganglion was decreased in acute migraine rats, suggesting that P2X7 receptor may play a role in migraine. The expression of c-fos and CGRP in spinal trigeminal nucleus of migraine was further increased after intraperitoneal injection of BBG in acute migraine rats, suggesting that the P2X7 receptor of satellite glia might play a protective role in migraine.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R747.2;R-332

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